Aquaculture 426427 (2014) 105111

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Aquaculture
journal homepage: www.elsevier.com/locate/aqua-online

The size of biooc determines the nutritional composition and the

nitrogen recovery by aquaculture animals
Julie Ekasari a,b, Deasy Angela a, Setiyo Hadi Waluyo c, Tauq Bachtiar c, Enang Harris Surawidjaja a,
Peter Bossier b, Peter De Schryver b,
a
b
c

Department of Aquaculture, Faculty of Fisheries and Marine Science, Bogor Agricultural University, Indonesia
Laboratory of Aquaculture and Artemia Reference Center, Ghent University, Belgium
Center for the Application of Isotopes and Radiation Technology, National Nuclear Energy Agency, Indonesia

a r t i c l e

i n f o

Article history:
Received 6 August 2013
Received in revised form 8 November 2013
Accepted 15 January 2014
Available online 29 January 2014
Keywords:
Biooc
Size distribution
Nitrogen recovery
Amino acid

a b s t r a c t
The effect of biooc size on the nutritional composition of the ocs and the nitrogen utilization by white shrimp
(Litopenaeus vannamei), red tilapia (Oreochromis niloticus) and mussels (Perna viridis) was investigated. Biooc
was collected from a shrimp culture unit and labeled with (15NcH4)2SO4. The ocs were sieved grouping them
into 4 different size classes (un-sieved, b 48 m, 48100 m, and N 100 m) and subsequently offered to shrimp,
red tilapia and mussels. The biooc class of N 100 m contained the highest levels of protein (27.8%) and lipid
(7.5%), whereas the biooc of b48 m seemed to be richest in essential amino acids. Based on the Essential
Amino Acid Index (EAAI), biooc produced in this study can be considered as a good quality protein source for
shrimp (0.930.97) and a useful protein source for tilapia (0.830.90) and mussel (0.810.88). The total amount
of nitrogen that could be derived from biooc was the highest when the biooc was larger than 100 m, i.e. 4.06
g N/kg shrimp, 3.79 g N/kg tilapia, and 1.17 g N/kg mussel, respectively. The nitrogen recovery from the biooc,
however, was the highest when the oc was b 48 m. Overall, this study showed that biooc consumption by
shrimp, red tilapia and mussels occurs irrespective of oc size but that oc size can play an important role in
the quality of biooc in terms of nutritional composition and nitrogen retention by the animals.
2014 Elsevier B.V. All rights reserved.

1. Introduction
Biooc technology has been widely studied and applied in aquaculture. This system applies the principle of assimilation of excreted dissolved nitrogen by heterotrophic bacteria by managing the C/N ratio
in the water (Avnimelech, 1999). This heterotrophic bacterial biomass
further forms aggregates (biooc) comprising not only the bacteria itself but also other microorganisms such as microalgae and zooplankton,
as well as trapped (in)organic particles or solids (De Schryver et al.,
2008). Biooc technology is not only an adequate approach in maintaining water quality in the aquaculture system but it also generates biomass that can contribute as a protein source for the cultured
organisms in situ (Avnimelech, 2009; Crab et al., 2010) or can be harvested for use as a feed ingredient (Kuhn et al., 2009, 2010). Hence,
the use of biooc as a food source may imply a decrease in the requirement of formulated feed protein (Xu et al., 2012) and also improve nitrogen utilization efciency by the cultured animals (Avnimelech,
2006). In order to evaluate the use of biooc as a food source, general
criteria of aquaculture feed can therefore be applied, i.e. the size of

Corresponding author at: Rozier 44, B-9000 Gent, Belgium. Tel.: + 32 9 264 37 54;
fax: + 32 9 264 42 93.
E-mail address: peter.deschryver@ugent.be (P. De Schryver).
http://dx.doi.org/10.1016/j.aquaculture.2014.01.023
0044-8486 2014 Elsevier B.V. All rights reserved.

particles, attractiveness and palatability, digestibility, and nutritional

content (Tacon, 1987b).
Specically for in situ biooc utilization, particle size distribution of
the food may be of inuence on the efciency by which cultured animals with different feeding behavior (lter feeders, scavengers, etc.)
utilize the ocs as feed. Furthermore, in an aquaculture system with
biooc technology application, oc size may also relate to the uptake
potential of the biooc by the cultured organism, the digestibility of
the ocs as well as the nutritional value of the ocs (De Schryver
et al., 2008).
The protein, lipid, carbohydrate and vitamin C contained within the
biooc represent a considerable fraction of the nutritional requirements
of several aquaculture species (Crab et al., 2010; Tacon et al., 2002). This
has been supported by previous studies which showed that biooc supplementation into shrimp feed (Kuhn et al., 2010) or biooc offering as a
food source to pink shrimp resulted in comparable results in growth and
survival to the control feeds (Emerenciano et al., 2012). Others have
reported that the application of biooc technology improved the feed
conversion and protein retention indicating indirectly that the consumption of biooc contributes to the growth of the cultured organisms
(Avnimelech, 2009; Gao et al., 2012; Hari et al., 2004; Wasieliesky et al.,
2006; Xu et al., 2012). Further investigations, however, are still needed
on the amino acids, fatty acids and micronutrients such as vitamins and
mineral contents in biooc.

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J. Ekasari et al. / Aquaculture 426427 (2014) 105111

This study investigated the effects of biooc particle size on its consumption and nitrogen utilization by Pacic white shrimp (Litopenaeus
vannamei), red tilapia (Oreochromis niloticus) and mussel (Perna viridis).
For this purpose, the stable isotope 15N was used as a tracer. In addition,
we examined the relationship of the nitrogen retention from the biooc
by the tested animals with the essential amino acid proles for biooc of
different particle size.
2. Materials and methods
2.1. Biooc preparation, 15N enrichment, and multilevel ltration
The schematic summary of the overall experiment is presented in
Fig. 1. A biooc suspension was prepared in a 300 L ber tank lled
with seawater and stocked with 60 shrimp (5 g). The biooc production
unit as well as the experimental units (see below) were located indoors
at a light/dark regime of 12 h/12 h and light intensity of 450 lx. Feed pellets containing 40% protein (Chuen Sin, PT. Grobest Indomakmur,
Indonesia) were administered three times daily at a total level of 8% of
the shrimp biomass day1. To promote biooc formation, 25 g of molasses containing 44% of carbon (C) was added daily to obtain an estimated
C/N ratio of 20. The biooc preparation was performed for 3 weeks until
total suspended solids (TSS) reached a level of more than 500 mg L1.
The shrimp and part of the water were subsequently removed, whereas
the remaining 40 L of the shrimp culture medium containing the majority of the biooc was enriched with 15N. The addition of (15NH4)2SO4
(20% atom excess (a.e.) 15N) was performed at a 15N enrichment level
of 12.5% on total N in biooc biomass, coupled with the addition of molasses (Avnimelech and Kochba, 2009) at an estimated C/N ratio of 20.
The suspension was gently aerated for 2 days until total ammonia nitrogen (TAN) of the oc suspension was 0 mg L1.
Part of the enriched biooc suspension was diluted using fresh seawater to obtain an estimated TSS of 500 mg L1. This was considered
as the un-sieved biooc group. The remaining suspension was sieved
over 2 stacked nylon lters of 100 m and 48 m mesh size, respectively,
to obtain the N 100 m, 48100 m, and b48 m oc size groups. A subsample of the enriched oc suspension was brought on the sieves and
gently swirled to avoid inducing disintegration of the ocs until all suspension passed through. The ocs that did not pass the lters (N100 m
group and 48 m100 m group) were gently transferred to fresh seawater before the next volume of biooc suspension was sieved. Finally,
all oc size classes were resuspended in seawater at an estimated TSS
concentration of 500 mg L1 and aerated for 1 h resulting in 4 different
batches of ocs produced with different sizes: original un-sieved ocs,
ocs which passed through the 48 m sieve (b 48 m), ocs which
Preparation

Feed 40% CP

passed through the 100 m but not the 48 m sieve (48 m100 m),
and ocs which did not pass the 100 m sieve (N 100 m). Five samples
from each batch were collected for total suspended solid (TSS) measurement to determine initial TSS. In addition, duplicate samples of
each biooc suspension were analyzed in terms of particle size distribution, nutritional composition, total nitrogen content and 15N content. To
determine the mass fractions of the b 48 m, 48100 m, and N100 m
oc size groups in the un-sieved biooc group, 1 L of un-sieved biooc
suspension was fractioned into the respective oc size groups according
to the previously described procedure, and the dry weight of each size
group was determined after drying at 105 C for 4 h.
2.2. Feeding experiment
Following resuspension, each size class of 15N labeled biooc was
distributed into 20 plastic tanks of 2 L (80 tanks in total). From each experimental species (shrimp, sh, and mussels), one individual was
stocked per tank (5 replicates per oc size class). The remaining 5 replicated tanks per oc size class lled with only biooc suspensions
were used to observed possible biooc degradation during the experiment. Five additional replicate tanks per species were also prepared as
a control, lled with seawater without any biooc addition and stocked
with one individual. Aeration was provided in each tank to ensure a homogenous suspension of oc particles. The stocking sizes of the shrimp,
red tilapia, and mussels were 10.6 1.2 g, 9.6 1.2 g, and 10.5 1.2 g
(67 cm in length), respectively. The animals were adapted to the environmental conditions of the experiment, i.e. the temperature, light condition and salinity in the laboratory for 1 week prior to the start of the
feeding experiment. As for red tilapia, the adaptation to seawater salinity was performed three weeks before the experiment. Twenty-four
hours before the biooc feeding experiment, feeding with commercial
pellet was stopped to ensure that the animals would feed on the biooc
suspension. The animals were kept in the biooc suspensions for 4 days
during which no additional feed was provided.
2.3. Sampling of animals
The total N and 15N content in the test animals was determined after
the feeding experiment. For this purpose, each animal was transferred
to clean seawater and allowed for gut evacuation for 2 h, which in preliminary tests using 15N labeled biooc was shown adequate to obtain
constant 15N body values indicating complete emptying of the gut of
the different species. Subsequently, the animals were sacriced and
oven dried (70 C for 24 h) for further analyses.

Enrichment

External C, C/N 20

External C,
C/N 20

Multilevel filtration

Feeding

(15NH4)2SO4
enrichment
Un-sieved
< 48 m
48 100 m
> 100 m

biofloc

White shrimp culture tank

200 shrimp/m3

21 days biofloc preparation

Biofloc bioreactor
Sampling:
Floc size distribution,
protein, lipid and amino
acids composition, TSS,
15N content

Sampling:
15N content

4 days feeding experiment

Fig. 1. Schematic summary of the experimental procedure. CP = crude protein content, C = carbon, TSS = total suspended solids.

J. Ekasari et al. / Aquaculture 426427 (2014) 105111

2.4. Physical and chemical analyses

The size distribution of each sieved biooc fraction was measured
using a Coulter particle size distribution counter (Coulter LS 100Q
Laser Diffraction Particle Size Analyzer, Beckman Coulter, USA) at a
size range of 0.31000 m. The distribution of particle sizes is expressed
as the percentage of each size by volume yielding a distribution curve
where the total area under the curve represents the total biooc volume, which is 100%. The volume fraction of each biooc size class was
calculated according to the summation of biooc volume (% of total volume) for this range under the curve. Biooc proximate composition was
analyzed following the procedure described in Takeuchi (1988). Total
suspended solids and total ammonia nitrogen (TAN) were analyzed according to Standard Methods for Water Quality Analysis (APHA, 2005).
15
N stable isotope analyses were performed using mass spectrophotometry following the procedure of IAEA (2001).
The amino acid composition of the biooc was measured by a
professional laboratory (Sarawanti Indo Genetech, Indonesia) using
high performance liquid chromatography (HPLC). Briey, a biooc
sample (100 mg) was hydrolyzed with 5 mL 6 N HCl for 22 h at

107

100 C. The sample was subsequently diluted in 50 mL distilled water

and ltered through a 0.45 m lter. Into a 500 L aliquot, 40 L of internal standard (alpha amino butyric acid) and 460 L of distilled water
were subsequently added. Prior to HPLC injection (AccOtag column
3.9 150 mm, 37 C, AccQTag eluent A), pre-column derivatization
was performed using a AccQ-Tag reagent kit followed by incubation
for 1 h at 55 C. Validation of the method was performed by using a
standard amino acid mixture (Thermo Scientic Amino Acid Standard
H catalog no. NCI0180) and a reference protein sample (bovine serum
albumin, Sigma Aldrich catalog no. P3569), and amino acid recovery
was obtained in a range of 91.3%106.9%).
A selection of amino acids that is considered as essential for aquatic
animals in general was chosen as described by Tacon (1987) and
Babarro et al. (2011). The essential amino acid ratio (E/A) for each essential amino acid (EAA) was expressed as the percentage of this
amino acid on the total amount of essential amino acids measured
(Arai, 1981). The essential amino acid index (EAAI) was calculated according to Penaorida (1989) using the formula:
EAAI

p
9
aa1=AA1  aa2=AA2   aa9=AA9:

Fig. 2. Volumetric oc size distribution of a) the un-sieved biooc (mean particle diameter 73.83 54.30 m), b) the biooc of the b48 m fraction (mean particle diameter 26.84
14.70 m), c) the biooc of the 48100 m fraction (mean particle diameter 50.75 27.60 m), and d) the biooc of the N100 m fraction (mean particle diameter 119.4
71.80 m). The area under the curve represents the total oc volume, which is 100%.

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J. Ekasari et al. / Aquaculture 426427 (2014) 105111

be attributed to the density of lamentous bacteria in the oc

(Barbusiski and Kocielniak,1995).

Table 1
Nitrogen, protein and lipid content of biooc of different size classes.
Biooc size class

Nitrogen content
(% on dry weight)

Protein content
(% on dry weight)

Lipid content
(% on dry weight)

Un-sieved
b48 m
48100 m
N100 m

4.0
2.8
3.7
4.5

25.0
17.2
23.4
27.8

7.2
6.7
6.0
7.5

with aa being the essential amino acid ratio (E/A) in the biooc, AA
being the E/A ratio in the animal body, and 1, 2, 3,, 9 being each of
the essential amino acids.
2.5. Statistical analyses
Homoscedasticity and normality of biooc uptake, total N uptake
and N recovery were assessed using Levene's test and a Kolmogorov
Smirnov test, respectively. It was found that the variances of these variables were not equal, therefore a non-parametric test (KruskalWallis
test) was applied, followed by a MannWhitney U test (P b 0.05).
3. Results and discussion
3.1. Biooc size distribution
The particle size distribution in relation to the volume of biooc before sieving is presented in Fig. 2a. The smallest (b 48 m) biooc size
was dominating as 44.8% of the total biooc volume consisted of b48
m oc particle. The 48100 m and N100 m oc fractions represented
26.0% and 29.2%, respectively, of the total volume. The dominance of
ocs of b48 m in the un-sieved biooc was also supported by the
mass fractions of the biooc suspension before sieving as this was 53%
(1.86 g) for the b48 m oc fraction, 12% (0.42 g) for the 48100 m
oc fraction, and 36% (1.25 g) for the N 100 m oc fraction, respectively.
Expressed as volume/weight (%/%) ratio, this shows that the ocs of 48
100 m were less dense (2.20) as compared to the b48 m ocs (0.85)
and the N 100 m ocs (0.82). The low density of oc aggregates may

3.2. Biooc protein, lipid and essential amino acid composition

Interestingly, biooc of different sizes was characterized by a different nutritional composition. The protein content of the biooc increased
according to the particle size (Table 1). The N100 m oc size group also
contained the highest lipid content, whereas the lowest lipid content
was observed in the 48100 m biooc. The high protein and lipid content of the ocs of N 100 m might be attributed to the concentration of
extracellular polymeric substances (EPS) which account for 8095% of
the organic matter in the ocs (Wilen et al., 2008). Protein content in
the diet is an important factor for the nutrition of most aquatic organisms. Most of the aquaculture species require protein at a range of 20
50% in their diet (Tacon, 1987a).
The amino acid composition of the biooc differed amongst biooc
size groups (Fig. 3). In general, biooc seems to be rich in valine, lysine,
leucine, phenylalanine and threonine but decient in methionine. Furthermore, though not essential, there was no cysteine found in any of
the biooc group. Protein quality of a diet for aquaculture species is
determined by the amino acid composition of the protein and the bioavailability of the amino acids present. According to the nutritional requirement and the capability of amino acid synthesis by an organism,
an amino acid is classied as nutritionally essential (indispensable) or
non essential (dispensable) for sh (Li et al., 2009). EAAs are those
that either cannot be synthesized or are inadequately synthesized de
novo by animals relative to their needs and consequently must be provided by the diet. As amino acids are building blocks for protein, an optimal synthesis of protein is therefore determined by the dietary amino
acid prole (Mente et al., 2002). It has been reported that the dietary
amino acid requirement of a particular aquatic organism is strongly dependent on its body amino acid composition (Tacon, 1987a).
Penaorida (1989) suggested the calculation of the EAAI to evaluate
the EAA prole in the diet relative to the EAA composition of the animal
and classied a diet to be of good quality with an EAAI of more than 0.9,
to be useful when it is in the range of 0.70.9, and to be inadequate
when it is less than 0.7. The EAAI values in Fig. 4 show that the EAA composition of the biooc of all sizes in general could meet the requirement

180
160

EAA Concentration (mol/g DW)

Arginine

140

Histidine
Isoleucine

120
Leucine

100

Lysine
Methionine (+
Cysteine)
Phenilalanine (+
Tyrosine)
Threonine

80
60

Valine

40
20
0

Un-sieved

< 48m

48-100m

>1

Floc particle size

Fig. 3. Essential amino acid pattern (mol g1 oc dry weight) of biooc of different size classes.

J. Ekasari et al. / Aquaculture 426427 (2014) 105111

109

oc size groups in this study. The gure also shows the situation for
each of the EAAs individually as it relates the E/A ratio of the biooc
(what is available) with the E/A of the animal (what is required according to literature data). The diagonal line represents the situation where
the E/A ratio of the biooc equals that of the animal and is the ideal case
(Tacon, 1987a). Data points that are situated below the diagonal line
then indicate an excess of that EAA in amino acid prole of the biooc,

of the aquatic species tested with a range of EAAI of 0.810.97. Based on

this index, the biooc produced in this study can be considered as a
good quality protein source for the shrimp and a useful protein source
for both tilapia and mussel. The gure also shows that ocs with a particle diameter of more than 100 m consistently showed the highest
EAAI for the three species tested, which suggests that the N100 m
oc size group was higher in quality in terms of EAAs than the other

22
20

Arginine

Phenylalanine

E/A ratio in shrimp (%)

18
16
Leucine

14

Un-sieved, EAAI=0.96

12

Lysine

10

< 48m, EAAI=0.93

Valine
Threonine

Isoleucine

48 -100m, EAAI=0.95

Methionine

Histidine

>100m, EAAI=0.97

2
0

10

12

14

16

18

20

22

E/A ratio in floc (%)

22

20

E/A ratio in tilapia (%)

18

Lysine

16
Phenylalanine

Leucine

14
12

Arginine

10
Methionine

8
6

Threonine
Valine
Isoleucine

Un-sieved, EAAI=0.90

Histidine

48 -100m, EAAI=0.88

< 48m, EAAI=0.83

4
>100m, EAAI=0.90

2
0
0

10

12

14

16

18

20

22

E/A ratio in floc (%)

Isoleucine

20

E/A ratio in mussel (%)

Histidine

18
16

Methionine

14

Un-sieved, EAAI=0.88

Lysine

12

< 48m, EAAI=0.81

10

Leucine
Arginine

Phenylalanine
Threonine

48 -100m, EAAI=0.85
>100m, EAAI=0.88

Valine

4
2
0

10

12

14

16

18

20

22

E/A ratio in floc (%)

Fig. 4. Relationship between the E/A ratio in ocs of different size classes with a) the E/A ratio in shrimp (shrimp whole body E/A ratio data from Tacon et al. (2002)); b) the E/A ratio in
tilapia (tilapia whole body E/A ratio data from Clement and Lovell (1994)); and c) the E/A ratio in mussels (mussel meat E/A ratio data from Babarro et al. (2011)). The 45 line represents
the situation where the E/A ratio in the ocs and in the animal is in perfect balance. Above the line, there is a shortage in the ocs in the essential amino acid under consideration whereas
below the line there is an excess. The EAAI for each oc size is indicated in the gure.

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J. Ekasari et al. / Aquaculture 426427 (2014) 105111

Table 2
Calculation of the biooc uptake by the animals during the experimental period.

ratio was considerably different from the mussel esh E/A ratio and it
was found to be decient in isoleucine, histidine and methionine.

Floc size
Un-sieved

b48 m

48100 m

N100 m

Initial biooc TSS (g/L)

0.41 0.09

0.20 0.08

0.48 0.08

0.58 0.03

Biooc TSS after 4 days (g/L)

Shrimp
0.25 0.03
Tilapia
0.0 0.0
Mussel
0.16 0.04

0.01 0.01
0.0 0.0
0.01 0.01

0.15 0.11
0.04 0.05
0.26 0.07

0.42 0.03
0.04 0.06
0.35 0.08

Biooc TSS reduction (g/L)

Shrimp
0.16 0.03
Tilapia
0.40 0.00
Mussel
0.24 0.04

0.19 0.02
0.20 0.00
0.18 0.02

0.33 0.12
0.44 0.06
0.22 0.10

0.17 0.03
0.54 0.09
0.23 0.08

Animal wet weight (g)

Shrimp
11.5 1.3
Tilapia
9.0 0.7
Mussel
10.2 1.1

10.4 1.5
10.0 1.0
10.2 1.1

10.6 1.3
9.4 0.9
10.3 1.2

9.8 0.6
9.3 0.9
10.4 1.5

Biooc uptake (g TSS/kg animal wet weight)**

Shrimp
27.9 6.5
36.5 7.2***
Tilapia 88.9 6.4ab*** 39.3 3.9a***
Mussel
47.9 4.3
36.0 4.8***

64.2 22.6
93.2 11.2ab***
57.2 22.4

34.3 7.6
117.3 24.9b***
41.6 5.5*

Different superscript letters in the same row indicate signicant differences (P b 0.05).
*n = 3 (due to 40% of mortality in mussel fed with N100 m size oc).
**Biooc uptake was calculated by the formula: Biooc TSS reduction (g/L) 2 L tank volume
1000 / animal wet weight (g).
***Minimal value () because biooc suspension was completely consumed after 4 days
of feeding experiment.

whereas data points situated above the diagonal line indicate a shortage. In this respect, it can be seen that there were some EAAs lacking
in each of the oc size classes depending on the species under consideration. For the shrimp, the biooc lacked mainly arginine and to a lower
degree leucine and methionine whereas for tilapia the ocs were relatively decient in methionine, arginine and lysine (Fig. 4a and b). The
requirement for EAAs of mussels seems to be different from shrimp
and tilapia (Fig. 4c). It can be seen that for mussels, the biooc E/A

3.3. Biooc consumption

The calculated consumption of the ocs by the tested animals is
given in Table 2. All animal species completely consumed the b48 m
ocs, and therefore the calculated TSS uptake values for the b 48 m
ocs should be considered as minimum values and would thus most
probably have been higher in the case of more biooc availability.
Though the differences were not signicant (P b 0.05), the highest oc
consumption by shrimp and mussels was observed for the ocs of 48
100 m (64 and 57 g TSS/kg biomass, respectively). Tilapia completely
consumed the biooc irrespective of the oc size. The oc consumption
of tilapia was in the range of 39 to 117 g TSS/kg tilapia, but since tilapia
was observed to completely consume all the ocs on the second day of
experiment, the actual oc consumption could be higher than these
values. Furthermore, it is also important to note that the calculated
TSS uptake may have been a slight underestimation of the actual oc
consumption as there was the possibility of minor oc formation during
the experiment due to nutrient recycling. This is suggested by the observation in the biooc control tanks where an average TSS increase of 12%
occurred (data not shown).
The data show that tilapia can harvest all the ocs regardless of size,
and this could be attributed to the nature of tilapia allowing it, as a lter
feeder, to harvest also the small-sized biooc particles. As shrimp have a
nibbling feeding behavior, it was expected that the shrimp at 10 g size
would prefer to consume bigger oc. The results however showed
that shrimp at this size also consumed small oc particles. Surprisingly,
mussels appear to also consume larger oc sizes. However, it is important to note that feeding with N 100 m ocs resulted in mortality of
part of the mussels (40%). This is in accordance with a previous report
(Tantanasarit et al., 2013) in which it was noted that mussel gills
would likely become clogged and ltration rate reduced, when exposed
to a solution with too many high-sized particles. This evidently implies
that the application of biooc technology in mussel culture will be limited to smaller particle sizes of biooc.

Table 3
Nitrogen content of the ocs, and nitrogen recovery by shrimp, tilapia and mussel from ocs of different sizes. Calculation of the nitrogen uptake by the animals from the biooc is
performed by means of the stable isotopic 15N tracer data (15NH4)2SO4.
Floc size
Un-sieved

b48 m

48100 m

N100 m

6.6
32.8

3.1
11.0

8.6
35.7

2.2
51.6

N % a.e.
Shrimp
Tilapia
Mussel

0.18 0.09
0.39 0.15
0.60 0.14

0.30 0.17
0.38 0.15
0.28 0.11

0.59 0.37
0.49 0.14
0.34 0.23

0.43 0.09
0.47 0.17
0.31 0.04

Total N in animals (mg N/animal)

Shrimp
Tilapia
Mussel

242 21
170 3
57 4

225 40
187 19
60 7

236 16
176 17
59 5

198 27
159 17
93 28

Total N uptake from biooc (mg N/animal)

Shrimp
Tilapia
Mussel

74
10 4
51

11 1
27 3
52

19 9
10 4
64

40 12
34 8
13 2

N recovery (%)
Shrimp
Tilapia
Mussel

21 12
32 12ab
16 4

53 26
29 10a
18 10

78 23
66 15ab
26 4

15

N % a.e. in ocs
Total N available in biooc (mg)
15

95 5
245 29b
49 18

Calculated by the formula: Initial biooc TSS (g/L) 2 L tank volume nitrogen content of biooc (%).
n = 3 (due to 40% of mortality in mussel fed with N100 m size oc).
Calculated by the formula (IAEA, 2001): 15N in animal (% a.e.) Total N in animal (mg N) / 15N in biooc (% a.e.).
Different superscript letters in the same row indicate signicant differences (P b 0.05). N recovery was calculated by the formula: (Total N derived from biooc / Total N available in
biooc) 100%.

J. Ekasari et al. / Aquaculture 426427 (2014) 105111

3.4. Nitrogen recovery from the ocs

The nitrogen in the animals that could be derived from the biooc
during the feeding experiment is given in Table 3. The nitrogen uptake
from biooc with a particle size of N100 m was found to be the highest
regardless of the animal species under consideration. Shrimp and mussel seemed to take up the least N from un-sieved ocs (7 mg N/shrimp
and 5 mg N/mussel). Tilapia showed the least N uptake from un-sieved
and the 48100 m ocs (10 mg N/tilapia). Regardless of the N uptake
values, consumption of the b48 m biooc leads to the highest N recovery from the biooc by all the animals tested, which may suggest that
this particular oc size was more digestible and better utilized. A more
than 100% value was noticed for the b48 m ocs consumed by tilapia
(245%) which was also signicantly higher than those of other oc
size groups. This value is not possible and is likely due to an inconsistency in one of the analyses, although it cannot be deducted from the data
in which value is erroneous. The trend for the N recovery does, however,
not change as the result of this value. Both shrimp and tilapia showed
a considerably higher N recovery from the biooc than mussel, irrespective of the oc size.
Bureau (2004) noted that the main factors affecting nitrogen retention by sh are amino acid composition (the concentration and prole),
and the balance between digestible protein and digestible energy.
Therefore, N recovery results may indicate that from a nutritional
point of view, biooc of b48 m and N 100 m particle sizes could
meet the requirement of the tested animals. This is supported by the
amino acid prole that is mentioned earlier. Although the protein content of b48 m oc was the least amongst other oc size groups, the
concentration of essential amino acids in this particular oc size was
generally the highest. The N 100 m oc class was also superior with regard to protein and lipid content as well as the essential amino acid balance represented by EAAI value.
4. Conclusion
Shrimp, tilapia and mussel were able to consume and retain N from
biooc at all particle sizes tested. Caution however must be taken in providing N100 m ocs for mussel. Overall results show that ocs with
particle size of N 100 m and b 48 m are more favorable for the aquaculture species tested in this study as they contain a higher nutritional
value and show higher N recoveries. Therefore particle size seems to
play an important role in the quality of biooc and consequently affects
the capacity of N retention by each animal. The capability of shrimp, tilapia and mussel in utilizing biooc in this study may also be used as a
basic information to develop an integrated multi-trophic-biooc system
in which managing suspended solids in BFT systems and enhancing nutrient utilization efciency for a more sustainable and environmentally
friendly aquaculture practices will be important.
Acknowledgment
This research was nancially supported by the Flemish Interuniversity CouncilUniversity Development Cooperation (VLIR). The rst
author would like to thank Dr. Muhammad Agus Suprayudi and
Dr. Widanarni for their assistance in data interpretation. P. De Schryver
is supported as a post-doctoral research fellow by the Fund for Scientic
Research (FWO) in Flanders (Belgium).
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